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[大鼠供肝冷保存及再灌注损伤过程中肝细胞凋亡及凋亡调节基因的表达]

[Hepatocyte apoptosis and expression of apoptosis-regulating genes during cold preservation and reperfusion injury in rat donor liver].

作者信息

Wang Yu, Kan He-ping, Jiang Xiao-qing

机构信息

Department of Hepatobiliary Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2010 Jul;30(7):1699-702.

Abstract

OBJECTIVE

To investigate the apoptosis of hepatocytes and the expression of apoptosis-regulating genes during the donor liver ischemia and reperfusion injury in rat orthotopic liver transplantation (OLT).

METHODS

Seventy-two male SD rats were randomly divided into sham operation group and transplantation group. Using Ringer's lactate solution as the perfusing and preserving solution, the grafts were preserved for 4 h before orthotopic transplantation. At 1, 6 and 24 h after the reperfusion, the recipients were sacrificed, and the serum ALT and AST levels were measured; the changes of hepatocyte apoptosis was detected by TUNEL assay, and the protein expressions of the apoptosis-regulating genes were measured by flow cytometry.

RESULTS

Serum ALT and AST levels were significantly higher in transplantation group than in the control group after reperfusion. In comparison with the control group, the rats in the transplantation group showed significantly increased apoptosis index in the livers, lowered Bcl-2 levels and increased FasL levels after the transplantation, especially at 6 h after liver reperfusion (P<0.01).

CONCLUSION

The donor liver ischemia and reperfusion injury can promote hepatocyte apoptosis, which may be related with the high expression of Bcl-2 gene and low expression of FasL after reperfusion injury in rats with orthotopic liver transplantation.

摘要

目的

探讨大鼠原位肝移植(OLT)供肝缺血再灌注损伤过程中肝细胞凋亡及凋亡调控基因的表达情况。

方法

将72只雄性SD大鼠随机分为假手术组和移植组。以乳酸林格液作为灌注和保存液,在原位移植前将移植物保存4小时。再灌注后1、6和24小时,处死受体,检测血清ALT和AST水平;采用TUNEL法检测肝细胞凋亡变化,通过流式细胞术检测凋亡调控基因的蛋白表达。

结果

再灌注后移植组血清ALT和AST水平显著高于对照组。与对照组相比,移植组大鼠移植后肝脏凋亡指数显著升高,Bcl-2水平降低,FasL水平升高,尤其是在肝脏再灌注后6小时(P<0.01)。

结论

供肝缺血再灌注损伤可促进肝细胞凋亡,这可能与原位肝移植大鼠再灌注损伤后Bcl-2基因高表达和FasL低表达有关。

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